ArticleBMC immunology2007
Successful downstream application of the Paxgene Blood RNA system from small blood samples in paediatric patients for quantitative PCR analysis.
Article in BMC immunology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
24 citing papers in PubMed, 56 citations in OpenAlex.
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- Transcriptional Biomarkers for Treatment Monitoring of Pulmonary Drug-Resistant Tuberculosis: Protocol for a Prospective Observational Study in Indonesia.Tropical medicine and infectious disease · 2022Article
- Validation of reference genes for whole blood gene expression analysis in cord blood of preterm and full-term neonates and peripheral blood of healthy adults.BMC genomics · 2021Article
- PICALM mRNA Expression in the Blood of Patients with Neurodegenerative Diseases and Geriatric Depression.Journal of Alzheimer's disease : JAD · 2021Article
- Cytokine and Gene Expression Profiling in Patients with HFE-Associated Hereditary Hemochromatosis according to Genetic Profile.Acta haematologica · 2021Article
- ABCA7 Gene Expression and Genetic Association Study in Schizophrenia.Neuropsychiatric disease and treatment · 2020Article
- Article
- Gene expression profiling of whole blood: A comparative assessment of RNA-stabilizing collection methods.PloS one · 2019Article
- Single CpG hypermethylation, allele methylation errors, and decreased expression of multiple tumor suppressor genes in normal body cells of mutation-negative early-onset and high-risk breast cancer patients.International journal of cancer · 2018Article
- Peripheral blood RNA gene expression in children with pneumococcal meningitis: a prospective case-control study.BMJ paediatrics open · 2017Article
- Evaluation of mRNA Biomarkers to Identify Risk of Hospital Acquired Infections in Children Admitted to Paediatric Intensive Care Unit.PloS one · 2016Article
- Postnatal Age Is a Critical Determinant of the Neonatal Host Response to Sepsis.Molecular medicine (Cambridge, Mass.) · 2015Article
- TREM2 mRNA Expression in Leukocytes Is Increased in Alzheimer's Disease and Schizophrenia.PloS one · 2015Article
- Epipolymorphisms within lipoprotein genes contribute independently to plasma lipid levels in familial hypercholesterolemia.Epigenetics · 2014Article
- Evaluation of a solid matrix for collection and ambient storage of RNA from whole blood.BMC clinical pathology · 2014Article
- Comparison of three different kits for extraction of high-quality RNA from frozen blood.SpringerPlus · 2014Article
- PAX3 in neuroblastoma: oncogenic potential, chemosensitivity and signalling pathways.Journal of cellular and molecular medicine · 2014Article
- Cellular defense system gene expression profiling of human whole blood: opportunities to predict health benefits in response to diet.Advances in nutrition (Bethesda, Md.) · 2012Review
- Efficient recovery of whole blood RNA--a comparison of commercial RNA extraction protocols for high-throughput applications in wildlife species.BMC biotechnology · 2012Article
- Systems immunology of human malaria.Trends in parasitology · 2012Review
Corrections and comments
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Authors and funding
8 authors at 3 institutions in 3 countries.
Funding
Abstract
backgroundThe challenge of gene expression studies is to reliably quantify levels of transcripts, but this is hindered by a number of factors including sample availability, handling and storage. The PAXgene Blood RNA System includes a stabilizing additive in a plastic evacuated tube, but requires 2.5 mL blood, which makes routine implementation impractical for paediatric use. The aim of this study was to modify the PAXgene Blood RNA System kit protocol for application to small, sick children, without compromising RNA integrity, and subsequently to perform quantitative analysis of ICAM and interleukin-6 gene expression.Aliquots of 0.86 mL PAXgene reagent were put into microtubes and 0.3 mL whole blood added to maintain the same recommended proportions as in the PAXgene evacuated tube system. RNA quality was assessed using the Agilent BioAnalyser 2100 and an in-house TaqMan assay which measures GAPDH transcript integrity by determining 3' to 5' ratios. qPCR analysis was performed on an additional panel of 7 housekeeping genes. Three reference genes (HPRT1, YWHAZ and GAPDH) were identified using the GeNORM algorithm, which were subsequently used to normalising target gene expression levels. ICAM-1 and IL-6 gene expression were measured in 87 Malawian children with invasive pneumococcal disease.
resultsTotal RNA yield was between 1,114 and 2,950 ng and the BioAnalyser 2100 demonstrated discernible 18s and 28s bands. The cycle threshold values obtained for the seven housekeeping genes were between 15 and 30 and showed good consistency. Median relative ICAM and IL-6 gene expression were significantly reduced in non-survivors compared to survivors (ICAM: 3.56 vs 4.41, p = 0.04, and IL-6: 2.16 vs 6.73, p = 0.02).
conclusionWe have successfully modified the PAXgene blood collection system for use in small children and demonstrated preservation of RNA integrity and successful quantitative real-time PCR analysis.
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